Peptidases of Trichobilharzia regenti (Schistosomatidae) and its molluscan host Radix peregra S. Lat. (Lymnaeidae): construction and screening of cDNA library from intramolluscan stages of the parasite

Trichobilharzia regenti is a neurotropic bird schistosome,causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained.The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue.Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.     

Dietary Effects on Sperm Quality of Litopenaeus vannamei Broodstock

A 56‐d feeding trial was conducted to investigate the effect of diet on sperm quality of Pacific white shrimp Litopenaeus vannamei broodstock. Dietary treatments consisted of a combination of 75% dry maturation diet and 25% fresh‐frozen squid (dry‐weight basis). Supplemental nutrients of the maturation diet were selectively deleted and replaced with wheat starch to produce the following treatments: 1) 75% basal maturation diet plus 25% squid (control); 2) 75% maturation diet without supplemental vitamins plus 25% squid; 3) 75% maturation diet without supplemental cholesterol and phospholipids plus 25% squid; 4) 75% maturation diet without supplemental astaxanthin plus 25% squid; and 5) a fresh diet composed of 60% squid and 40% Maine bloodworms. Shrimp fed the control diet and the diet without supplemental astaxanthin had significantly higher mean (± SEM) change in sperm count (4.6 ± 3.2 million sperm cells and 2.9 ± 2.5 million sperm cells, respectively), with respect to baseline (8.7 ± 1.0, 6.4 ± 1.0, 9.0 ± 1.3, 6.6 ± 0.7, and 6.0 ± 0.8 million sperm cells for treatments 1, 2, 3, 4, and 5, respectively), than shrimp fed the diet without supplemental vitamins (‐1.7 ± 2.6 million sperm cells), but not significantly higher than those of shrimp receiving the diets without supplemental cholesterol‐phospholipids (1.2 ± 2.5 million sperm cells) and the fresh diet (1.3 ± 1.6 million sperm cells). Dietary deficiencies also were reflected in weight gain of shrimp fed the diet without supplemental vitamins (‐2.0 g) and the fresh diet (‐0.8 g). which were significantly lower than weight gain of shrimp fed the control diet (1.1 g) and the diet without supplemental cholesterol‐phospholipids (0.8 g). No significant differences were detected among treatments for percentage of abnormal sperm and survival data. Results demonstrated a significant effect of diet on reproductive quality of male L. vannamei and indicated that the typical combination of fresh‐food organisms used is not nutritionally optimal for male broodstock.     

Single immunization with an inactivated vaccine protects sheep from Schmallenberg virus infection

The arthropod-borne Schmallenberg virus (SBV), family Orthobunyaviridae, emerged in Europe in 2011. SBV is associated with a mild disease in adult ruminants but fetal malformation after an infection during a critical phase of pregnancy. A number of inactivated vaccines have been developed; their efficacy after two injections was demonstrated. To make the vaccination of sheep more efficient and economic the effect of a single immunization with one of these vaccines was investigated in the present study. Five vaccinated sheep and five additional control sheep were inoculated with SBV three weeks after vaccination and the results of a competitive ELISA, a standard microneutralization test and an SBV-specific real-time RT-PCR confirmed vaccine efficacy by demonstrating complete inhibition of viral replication in immunized animals.     

Panagrellus redivivus mass produced on solid media as live food for Litopenaeus vannamei larvae

The free‐living soil nematode Panagrellus redivivus is well known to be an excellent food source for first feeding fish larvae. It represents an alternative to the highly expensive Artemia, which is commonly used. The lack of a proper method for mass production of P. redivivus has prevented its wider use in commercial hatcheries. A new cultivation method allows the production of a sufficient quantity of nematodes to deliver a standardized and permanently available live food of high quality, throughout the larval rearing period. In two experiments – carried out at the Centro de Investigación en Alimentación y Desarrollo, Mexico – several feeding regimes were established to prove the quality of the mass produced P. redivivus for larvae of Litopenaeus vannamei, the Pacific white shrimp. Two different nematode treatments were compared with a no‐feed group and a control group that was fed with Artemia. All treatments had an additional algal co‐feed and were run in five replicates. Panagrellus redivivus was cultured on two different media (wheat/corn flour and oat flour) to compare these for their suitability as high‐quality live food for the larvae. Shrimp fed nematodes grown on wheat/corn medium reached the postlarval stage earlier than those from other treatments. The nematode treatments showed promising results; however, further research is needed on the development of improved culture media or enrichment methods to further increase the nutritional value of P. redivivus.     

Protein metabolism during sexual maturation in female Atlantic salmon (Salmo salar L)

Body composition and fractional rates of protein synthesis (percentage of the protein mass synthesized per day) were determined in female Atlantic salmon returning to the River Tay, Scotland in July and in October after a 95 day period without food, during which time the animals became sexually mature. During the 95 day period of starvation/sexual maturation the ventricle and red muscle remained as a constant proportion of fresh weight whereas the liver, gill and ovary increased and the stomach and white muscle decreased. Fractional rates of protein synthesis increased markedly in the liver, stomach and ovary during the period of starvation/sexual maturation. In the gill, ventricle and white muscle fractional protein synthesis rates increased slightly or remained constant. From the estimated rates of protein loss or gain in the various tissues it is concluded that there is considerable protein turnover and repartitioning of amino acids during the period of starvation and sexual maturation. The absolute rate of protein synthesis rates in the ovary indicates that this tissue made the largest contribution to the energy and amino acid demands of the fish, whilst most of the amino acids required for maturation of the ovary were derived from white muscle, principally as the result of increased muscle protein degradation.     

Investigation of the Effects of Salinity and Dietary Protein Level on Growth and Survival of Pacific White Shrimp, Litopenaeus vannamei

It is presumed that in hypo‐ and hypersaline environments, shrimp’s requirements for some specific nutrients, such as protein, may differ from those known in the marine habitat; however, few investigations have been conducted in this area of study. In the present investigation, the effects of salinity and dietary protein level on the biological performance, tissue protein, and water content of Pacific white shrimp, Litopenaeus vannamei, were evaluated. In a 3 × 4 factorial experiment, juvenile shrimp with an average initial weight of 0.36 ± 0.02 g were exposed for 32 d to salinities of 2, 35, and 50 ppt and fed experimental diets with crude protein contents of 25, 30, 35, and 40%. A significant effect of salinity on growth of shrimp was detected, with the growth responses (final weight, weight gain) ranked in the order 2 ppt (3.87, 3.50 g) > 35 ppt (3.40, 3.04 g) > 50 ppt (2.84, 2.47 g). No effects of dietary protein level or an interaction between salinity and protein on growth of shrimp were observed under the experimental conditions of this study. Percent survival of shrimp fed the highest protein content (40%, survival of 74%) was, however, significantly lower than those of shrimp fed the other feeds (25, 30 and 35% protein, survival of 99, 91, and 94%, respectively), a result likely associated with the concentration of total ammonia nitrogen, which increased significantly at increasing protein levels. Final water content of whole shrimp was significantly lower in animals exposed to 50 ppt (70.8%) than in shrimp held at 2 (73.7%) and 35 ppt (72.3%). No effect of salinity, protein, or their interaction was observed on the protein content of whole shrimp. The results of the present study are in agreement with reports of superior and inferior growth of L. vannamei reared in hypo‐ and hypersaline environments, respectively, as compared to what is generally observed in seawater.     

MAGNETIC RESONANCE IMAGING CONTRAST ENHANCEMENT OF EXTRA‐OCULAR MUSCLES IN DOGS WITH NO CLINICAL EVIDENCE OF ORBITAL DISEASE

Enhancement of extra‐ocular muscles has been reported in cases of orbital pathology in both veterinary and medical magnetic resonance imaging. We have also observed this finding in the absence of orbital disease. The purpose of this retrospective study was to describe extra‐ocular muscle contrast enhancement characteristics in a group of dogs with no known orbital disease. Magnetic resonance images (MRI) from dogs with no clinical evidence of orbital disease and a reportedly normal MRI study were retrieved and reviewed. Contrast enhancement percentages of the medial, lateral, ventral, and dorsal rectus muscles were calculated based on signal‐to‐noise ratios that were in turn determined from hand‐traced regions of interest in precontrast, immediate postcontrast and 10‐min postcontrast scans. Comparison measurements were made in the pterygoid muscle. Contrast enhancement of the extra‐ocular muscles was observed in all patients (median contrast enhancement percentage 45.0%) and was greater than that of pterygoid muscle (median contrast enhancement percentage 22.7%). Enhancement of the extra‐ocular muscles persisted 10 min after contrast administration (median contrast enhancement percentage 43.4%). Findings indicated that MRI contrast enhancement of extra‐ocular muscles is likely normal in dogs.     

Recovery from neuromuscular block in dogs: restoration of spontaneous ventilation does not exclude residual blockade

ObjectiveTo evaluate if return of spontaneous ventilation to pre-relaxation values indicates complete recovery from neuromuscular blockade.Study designProspective, with each individual acting as its own control.AnimalsTen healthy adult female Beagle dogs weighing 6.2–9.4 kg.MethodsDogs were anesthetized with propofol, dexemedetomidine and isoflurane. Spontaneous ventilation was assessed by measuring end-tidal CO₂, expired tidal volume, peak inspiratory flow, respiratory rate and minute ventilation. Vecuronium 25 μg kg^?1 IV was administered and neuromuscular block was evaluated by measuring the train-of-four (TOF) ratio with acceleromyography in the hind limb. During spontaneous recovery from neuromuscular block, the TOF ratio when each ventilatory variable returned to baseline was recorded.ResultsThis dose of vecuronium produced moderate neuromuscular block in all dogs, with TOF ratio values of 0–18% at maximal block. Expired tidal volume, peak inspiratory flow and minute ventilation returned to pre-relaxation values when the median TOF ratio was ≤ 20%. The median TOF ratio was 42% when the end-tidal CO₂ returned to pre-relaxation values.Conclusions and clinical relevanceSignificant residual neuromuscular block could be measured at the hind limb with acceleromyography when ventilation had spontaneously returned to pre-vecuronium values. Monitoring spontaneous ventilation, including end-tidal CO₂, expired tidal volume, peak inspiratory flow or minute ventilation cannot be used as a surrogate for objective neuromuscular monitoring, and this practice may increase the risk of postoperative residual paralysis.